The goal of the first phase of GESTALT was to obtain transcriptomic, proteomic and epigenomic data from 11 purified immune cell subsets obtained from peripheral blood. Sample collection for this phase will be completed in October/November 2019. In the interim we have begun to analyze RNA, protein, DNA methylation and ATAC-Seq data from the first 54 samples. During FY19: 1)we continued sample collection to complete the objective of 100 donors across ages. 2)Transcriptomic analysis from the first 50 donors was used to define cell-specific patterns gene expression. Analysis of gene expression changes with age are ongoing. 3)Cell-specific patterns of DNA methylation were obtained and the relationship to cell-specific patterns of gene expression was determined. 4)Age-associated changes in DNA methylation were identified in each cell type. 5)Age-associated changes in chromatin structure was evaluated in monocytes using a small cohort of GESTALT samples; a larger scale analysis is ongoing. The results will be compared to similar studies carried pout with human CD8+ cells in the labs of Goronzy and Ucar. We also obtained ATAC-Seq data from nave and memory B cells; analysis is pending. 6)Flow cytometry panels were established to examine B, T and monocyte activation using total PBMC.